STANDARDISATION OF OCULAR

MICROMETER, DETERMINATION
OF MAGNIFICATION AND
CAMERA LUCIDA DIAGRAMS OF
RHIZOPUS AND SPIROGYRA.
SOHAM BISWAS
BIOLOGY PROJECT
DR. JYOTI PRASAD SARKAR
By
CERTIFICATE OF EFFORT
THIS IS TO CERTIFY THAT SOHAM BISWAS OF
BHOLANANDA NATIONAL VIDYALAYA HAS
HONESTLY COMPLETED THIS PROJECT UNDER MY
GUIDANCE AND SUPERVISION AND HAS SUBMITTED
THE PROJECT IN THE STIPULATED TIME .

DATE 15/1/2014
NAME OF STUDENT SOHAM BISWAS.
ROLL NO -
SCHOOL- BHOLANANDA NATIONAL VIDYALAYA.




SIGNATURE:
[ DR. JYOTI PRASAD SARKAR ]



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ACKNOWLEDGEMENT

I AM GRATEFULL TO DR. JYOTI PRASAD SARKAR, TEACHER
OF BIOLOGY FOR HIS NECESSARY GUIDANCE AND
SUPERVISION, WITHOUT WHICH THIS PROJECT WOULD NOT
HAVE SEEN THE LIGHT OF THE DAY.
I AM GRATEFULL TO MRS SWATI RUDRA, PRINCIPAL FOR
PROVIDING US THE LABORATORY TO CONDUCT THIS
PROJECT



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TABLE OF CONTENTS
INTRODUCTION ................................................................................ 3
UNITS OF MEASUREMENT IN MICROMETRY ........................... 5
STANDARISATION OF OCULAR MICROMETER ....................... 6
DETERMINATION OF STANDARISATION .................................. 8
MEASUREMENT OF SPORES, FILAMENTS.CELLS ................. 10
DETERMINATION OF MAGNIFICATION ................................... 11
CAMERA LUCIDA DIAGRAM OF RHIZOPUS AND
SPIROGYRA 12

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INTRODUCTION
Micrometry is the measurement of microorganisms. Since
microorganisms are small, the dimensions of these are usually expressed
in units smaller than millimeters. Microorganisms, such as viruses,
which are much smaller in size than bacteria, the nanometer and
angstrom, are used. A nanometer (nm) is one billionth (10) of a meter
(one- millionth of a millimeter). An angstrom is one-tenth of a
nanometer . Determination of size of a microorganism. is one of the
properties useful for identification in the laboratory. Since
microorganisms can be seen only under a microscope, their size can be
measured by equipping the microscope with an ocular micrometer,
which is then calibrated against a stage micrometer.
The ocular micrometer is simply a glass disc with etched lines on its
surface. It has 109 equally spaced divisions, marked 0-10 (see Fig. 1.9).
The distance between the graduations of an ocular micrometer does not
have any standard value and varies depending on the objective used, the
latter determines the size of the field, and this distance is found out by
calibrating it with a known scale, the stage micrometer.
Stage micrometer is a special glass slide having in its centre a known
distance one millimeter, which is encircled and mounted by a cover
glass .This one mm distance is etched into 100 equally spaced divisions.
Since there are 1000 micrometers in one millimeter. Thus one division
of stage micrometer equals to 10 0.01 mm. The distance of each stage
micrometers division becomes correspondingly Enlarged under high-
power and oil-immersion objectives of the microscope.
Ocular micrometer after putting inside the eye piece is calibrated by
superimposing the graduations of ocular micrometer over graduations of
the stage micrometer which is accomplished by rotating the ocular lens.

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By determining the number of ocular micrometer divisions coinciding
with the number of divisions on stage micrometer, the calibration factor
for one ocular division (O.D.) is calculated for the particular
combination of objective and ocular lens used by applying the formula:
- Number of divisions on stage micrometer
One ocular division (on lm) x 10
No. of divisions on ocular micrometer
After calibrating, an ocular micrometer can be used to determine the size
of an organism/structure of an organism in terms of length, breadth and
diameter by the formula:
Size (in m) = No. of ocular divisions x Calibration factor of the
objective used







OCULAR MICROMETER EYEPEICE


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UNITS OF MEASUREMENT IN MICROMETRY

MILLIMETER (mm)
1 mm= 0.001 m

MICROMETER (m)
1 m=0.001 mm
NANOMETER (nm)

1nm=0.001m




THE MICROMETER EYEPIECE





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STANDARISATION OF OCULAR MICROMETER

REQUIREMENTS:
COMPOUND MICROMETER
OCULAR MICROMETER
STAGE MICROMETER
PROCEDURE:
1. WE FIRST REMOVE THE EYE PIECE (OCULAR) LENS FROM OUR
MICROSCOPE, SCREW IT AND INSERT THE OCULAR MICROMETER
DISC ON THE CIRCULAR SHELF (METAL DIAPHRAGM) WITH THE
ENGRAVED SIDE DOWN INTO IT. SCREW THE EYE PIECE BACK AND
INSERT IT IN THE MICROSCOPE. WHEN WE OBSERVE OCULAR
MICROMETER DIVISIONS WILL BE SEEN IN SHARP FOCUS. AND
THERE WILL BE NO CHANGES IN LINES AND DISTANCES UNDER
DIFFERENT OBJECTIVES.

2. WE PLACE THE STAGE MICROMETER ON THE MICROSCOPE
STAGE AND BRING ITS SCALE IN THE MICROSCOPES FIELD
CENTER UNDER A SHARP FOCUS FIRST USING THE LOW-POWER
OBJECTIVE, AND THEREAFTER WITH HIGH-POWER OBJECTIVES

3. TURN THE OCULAR LENS UNTILL THE PARALLEL LINES OF
OCULAR MICROMETER BE PARALLEL WITH THOSE OF THE STAGE
MICROMETER.
4. MAKE THE LINES COINCIDE AT THE LEFT END AND FIND
ANOTHER LINE SET WHICH COINCIDES.

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5. NOW COUNT THE NUMBER OF DIVISIONS IN BOTH THE OCULAR
AND STAGE MICROMETER, BETWEEN THE TWO COINCIDING LINES.



6. TAKE ATLEAST 3 READINGS REPEATING STEPS 3-5.
7. REPEAT THE PROCEDURE (STEPS 3-6) WITH THE HIGH POWER
OBJECTIVE.


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DETERMINATION OF STANDARISATION
LOW POWER OBJECTIVE
STAGE DIVISION OCULAR DIVISION
5 7
3 4
4 5
AVERAGE 4 5.3
STANDARISATION = ) * 10 m
= (4%5.3)*10 m
I OCULARDIVISION= 7.547 m.








STAGE MICROMETER LINES

OCULAR MICROMETER LINES

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HIGH POWER OBJECTIVE
STAGE DIVISION OCULARDIVISION
2 5
3 8
1 3
2 5
AVERAGE 2 5.25
STANDARISATION = ) * 10 m
= (2%5.25)*10 m
= 3.809 m.
1 OCULARDIVISION = 3.809 m.

NOTE: IF YOU HAVE TO CHANGE MICROSCOPES THEN YOU HAVE
TO CALIBRATE EACH TIME AGAIN FOR EACH OF THE OBJECTIVE
LENSES YOU WILL BE USING. WHY? BECAUSE THE MAGNIFICATION
IS DIFFERENT ON DIFFERENT MICROSCOPES.


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MEASUREMENT OF SPORES, FILAMENTS.CELLS
UNDER HIGH POWER OBJECTIVE
NEUTROPHIL:
OCULAR DIVISION = 2
ACTUAL LENGTH = 2 *3.809=7.618m
UREDOSPORE:
LENGTH OF UREDOSPORE
OCULARDIVISION 8.5
ACTUAL LENGTH 32.37 m
BREADTH OF UREDOSPORE
OCULAR DIVISION- 4.
ACTUAL BREADTH 15.236m
SINGLE CELL OF SPIROGYRA
LENGTH:
OCULARDIVISION: 26
ACTUAL LENGTH 99.03 m
BREADTH 17
ACTUAL BREADTH-64.753m
ZYGOSPORE OF RHIZOPUS
DIAMETER
OCULARDIVISION 180
ACTUAL DIAMETER-685.6m.

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DETERMINATION OF MAGNIFICATION
REQUIREMENTS: CAMERA LUCIDA, STAGE MICROMETER, PENCIL,
SCALE.
LOW POWER OBJECTIVE
1 STAGE DIVISION= 10 m
100 m IS MAGNIFIED TO 1.6 cm
1.6 cm=16000 m.
MAGNIFICATION FACTOR = (1.6/100)*10000
= 160 X.




HIGH POWER OBJECTIVE
1 STAGE DIVISION= 10 m
100 m IS MAGNIFIED TO 6.9 cm
6.9 cm=69000 m.
MAGNIFICATION FACTOR = (6.9/100)*10000
= 690 X.




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CAMERA LUCIDA DIAGRAMS


1. DIAGRAM OF RHIZOPUS (160X)









2. DIAGRAM OF SINGLE CELL OF SPIROGYRA (160X)









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